Anti-Cytokeratin 6 antibody [EPR1602Y]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 6 antibody [EPR1602Y] (AB93279)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling Cytokeratin 6 with purified ab93279 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051 a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500).

Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 6 antibody [EPR1602Y] (AB93279)

Immunocytochemistry/Immunofluorescence analysis of A431 (Human epidermoid carcinoma cell line) cells labelling Cytokeratin 6 with purified ab93279 at 1/50. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077 an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291 a mouse anti-tubulin (1/500) and ab150120 an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500) were also used.

Control 1 : Primary antibody (1/100) and secondary antibody ab150120 an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500).

Control 2 : ab7291 (1/1000) and secondary antibody ab150077 an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 6 antibody [EPR1602Y] (AB93279)

IHC image of Cytokeratin 6 staining in a section of formalin-fixed paraffin-embedded normal human skin* performed on a Leica Biosystems BOND® RX instrument. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab93279, 0.5ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 6 antibody [EPR1602Y] (AB93279)

ICC/IF image of unpurified ab93279 stained A431 (Human epidermoid carcinoma cell line) cells.

The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab93279 1/100 dilution) overnight at +4°C. The secondary antibody (green) was ab96899 DyLight^® 488 goat anti-rabiit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor^® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 6 antibody [EPR1602Y] (AB93279)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human squamous cervical carcinoma tissue labelling Cytokeratin 6 with unpurified ab93279 at a dilution of 1/250.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-Cytokeratin 6 antibody [EPR1602Y] (AB93279)

Blocking buffer : 5% NFDM /TBST.

All lanes:

Western blot - Anti-Cytokeratin 6 antibody [EPR1602Y] (ab93279) at 1/1000 dilution

Lane 1:

N-GST tagged full-length recombinant human Cytokeratin 6A protein at 10 ng

Lane 2:

N-GST tagged full-length recombinant human Cytokeratin 5 protein at 10 ng

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 87 kDa

false

Exposure time: 180s

• WB

Unknown

Western blot - Anti-Cytokeratin 6 antibody [EPR1602Y] (AB93279)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Cytokeratin 6 antibody [EPR1602Y] (ab93279) at 1/2000 dilution

Lane 1:

Human skin tissue lysate at 20 µg

Lane 2:

Human tonsil tissue lysate at 20 µg

Secondary

All lanes:

Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 60 kDa

Observed band size: 56 kDa

false

• WB

Unknown

Western blot - Anti-Cytokeratin 6 antibody [EPR1602Y] (AB93279)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Cytokeratin 6 antibody [EPR1602Y] (ab93279) at 1/10000 dilution

All lanes:

HaCaT (Human keratinocyte cell line) cell lysate at 20 µg

Secondary

All lanes:

Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 54 kDa,60 kDa

Observed band size: 56 kDa

false

• WB

Unknown

Western blot - Anti-Cytokeratin 6 antibody [EPR1602Y] (AB93279)

All lanes:

Western blot - Anti-Cytokeratin 6 antibody [EPR1602Y] (ab93279) at 1/5000 dilution

All lanes:

A431 (Human epidermoid carcinoma cell line) cell lysate at 10 µg

Predicted band size: 60 kDa

false